JP2005316036A - Imaging apparatus and method and program to control illuminating light beam - Google Patents

Abstract

<P>PROBLEM TO BE SOLVED: To provide an imaging apparatus with which both pleasantness during observation and damage prevention of a sample such as color fading of the sample or the like are compatible by appropriately controlling the opening and the closing of the shutter of a microscope device in accordance with the operating mode of the imaging apparatus, and to provide a method and a program to control illuminating light beams executed by the imaging apparatus. <P>SOLUTION: The imaging apparatus picks up an observed image of the microscope device which is provided with a light source which emits illuminating light beams, an illuminating optical system which guides the illuminating light beams from the light source with which the sample is irradiated, a light shielding means which shields the illuminating light beams being guided to the sample and an image forming optical system that converges observation light beams outputted from the sample irradiated with the light beams, and forms an image. Moreover, the image pickup unit is provided with an imaging element which detects the observation light beams that are image formed by the image forming optical system, a display means which displays the image of the sample based on the observation light beams detected by the imaging element and a control means which controls the light shielding means corresponding to the operating mode being displayed by the display means. <P>COPYRIGHT: (C)2006,JPO&NCIPI

Description

  The present invention relates to an imaging device, an illumination light control method, and an illumination light control program, and in particular, a light source that emits illumination light, and an illumination optical system that guides the illumination light emitted from the light source to irradiate a sample. An observation image by a microscope apparatus comprising: a light shielding means for shielding the illumination light guided to the sample; and an imaging optical system for condensing and imaging the observation light output from the sample irradiated with the illumination light The present invention relates to an imaging device that captures images, an illumination light control method and an illumination light control program executed by the imaging device.

2. Description of the Related Art Conventionally, in fluorescence observation or fluorescence imaging using a microscope apparatus, in order to suppress damage such as discoloration of a sample due to illumination light, it is known to control the opening / closing of the illumination light shutter by a so-called trigger signal output from the imaging apparatus ( For example, see Patent Document 1.)
The configuration of a conventional microscope apparatus will be briefly described below.

  The illumination light emitted from the light source passes through the illumination lens and enters the half mirror. Of the illumination light incident on the half mirror, illumination light reflected by the half mirror passes through the objective lens and irradiates the sample placed on the stage. Here, assuming that the optical axis direction is the Z direction and two directions orthogonal to each other in a plane orthogonal to the Z direction are the X and Y directions, the stage can be moved in the XYZ directions by the stage drive system.

  The observation light reflected by the sample passes through the objective lens and enters the half mirror. The observation light transmitted through the half mirror passes through the imaging lens and forms an image on the image sensor. The output signal of the image sensor is converted into an image signal by the video signal processing circuit and then transferred to the monitor. As a result, an image of the sample is displayed on the monitor.

The shutter used in such a microscope apparatus receives a trigger signal so that the shutter opens during the exposure period when the image capturing apparatus acquires an image, and operates uniformly so that the shutter closes during other periods, and the illumination light is more than necessary. Is to prevent the sample from being irradiated (see, for example, Patent Document 2).
JP-A-7-199072 JP 2000-330033 A

However, in recent years, the operation method of imaging devices has been diversified, and various imaging methods such as display of moving images and acquisition of still images, or suppression of dark current noise of the imaging device due to exposure time and element temperature can be taken. It has become.
Therefore, it is impossible to achieve both observation comfort and prevention of damage to the sample by simply transmitting and blocking the illumination light according to the exposure period of the imaging device as in the known method. There was a problem.

For example, since the shutter is repeatedly transmitted / shielded during the exposure period of each frame even during moving image display, the shutter opening / closing sound continues to be emitted during observation by the moving image display of the imaging device, and the user hears annoying sound. There was a problem that comfort could not be secured.
Also, when acquiring multiple images of dark images and dark images to suppress dark noise, illumination light is irradiated onto the sample even during dark image shooting without the need to illuminate the sample. Therefore, there was a problem that damage to the sample could not be prevented.

  The present invention has been made in view of the above-described drawbacks of the prior art, and by appropriately controlling the opening and closing of the shutter of the microscope apparatus according to the operation mode of the imaging apparatus, the comfort of observation and the discoloration of the sample, etc. An object of the present invention is to provide an imaging apparatus capable of achieving both damage prevention, an illumination light control method and an illumination light control program executed by the imaging apparatus.

The present invention employs the following configuration in order to solve the above problems.
That is, according to one aspect of the present invention, an imaging apparatus according to the present invention includes a light source that emits illumination light, an illumination optical system for guiding the illumination light emitted from the light source to irradiate the sample, and the sample. An imaging apparatus that captures an observation image by a microscope apparatus, comprising: a light shielding unit that shields the illumination light to be guided; and an imaging optical system that focuses and images the observation light output from the sample irradiated with the illumination light. An imaging element for detecting observation light imaged by the imaging optical system, display means for displaying an image of the sample based on the observation light detected by the imaging element, and the display means. Control means for controlling the light shielding means corresponding to the operation mode to be displayed is provided.

  In the imaging apparatus of the present invention, the control unit controls the light shielding unit so that illumination light is always irradiated to the sample when the operation mode is the moving image display mode, and the operation mode is a still image. In the display mode, it is desirable to control the light shielding means so that illumination light is irradiated to the sample only during the exposure period.

  The image pickup apparatus of the present invention further includes a noise suppression unit that suppresses noise caused by a dark current of the image pickup device, and the control unit is in a mode in which the operation mode acquires a bright image. The light shielding means is controlled so that illumination light is irradiated to the sample only during the exposure period, and the illumination light to the sample during the exposure period is shielded when the operation mode is a mode for acquiring a dark image. Thus, it is desirable to control the light shielding means.

In the imaging apparatus of the present invention, the light source is an epi-illumination light source, the epi-illumination shutter is configured to illuminate the sample with the illumination light when the light-shielding means is open, and shield the illumination light by closing the light source. It is desirable to be.
In the imaging apparatus of the present invention, the light source is a transmissive illumination light source, the illuminating light is irradiated onto the sample when the light shielding means is open, and the transmissive shutter shields the illumination light when closed. It is desirable to be.

  According to another aspect of the present invention, the illumination light control method of the present invention includes a light source that emits illumination light, an illumination optical system that guides the illumination light emitted from the light source to irradiate the sample, and the above An observation image is picked up by a microscope apparatus having a light shielding means for shielding illumination light guided to the sample and an imaging optical system for condensing and imaging the observation light output from the sample irradiated with the illumination light. An illumination light control method executed by an imaging apparatus, wherein an operation mode is detected when an observation light imaged by the imaging optical system is detected and an image of the sample is displayed based on the detected observation light. The light shielding means is controlled corresponding to the above.

  In the illumination light control method according to the present invention, when the operation mode is the moving image display mode, the control unit controls the light shielding unit so that the illumination light is always applied to the sample, and the operation mode is stationary. In the image display mode, it is desirable to control the light shielding means so that the sample is irradiated with illumination light only during the exposure period.

  Further, the illumination light control method of the present invention further suppresses noise caused by the dark current of the image sensor, and the control is performed only during the exposure period when the operation mode is a mode for acquiring a bright image. The light shielding means is controlled so that illumination light is irradiated onto the sample, and the light shielding is performed so as to shield the illumination light on the sample during the exposure period when the operation mode is a mode for acquiring a dark image. It is desirable to control the means.

  Moreover, according to one aspect of the present invention, the illumination light control program of the present invention includes a light source that emits illumination light, an illumination optical system that guides the illumination light emitted from the light source to irradiate the sample, and the above An observation image is picked up by a microscope apparatus having a light shielding means for shielding illumination light guided to the sample and an imaging optical system for condensing and imaging the observation light output from the sample irradiated with the illumination light. An illumination light control program to be executed by an imaging apparatus, the procedure for detecting observation light imaged by the imaging optical system, and displaying an image of the sample based on the detected observation light In addition, a computer-executable illumination light control program for executing a procedure for controlling the light shielding unit corresponding to an operation mode.

  In the illumination light control program of the present invention, the control procedure controls the light shielding means so that the illumination light is always irradiated to the sample when the operation mode is the moving image display mode. However, in the still image display mode, it is desirable to control the light shielding means so that illumination light is irradiated to the sample only during the exposure period.

  The illumination light control program according to the present invention further includes a procedure for suppressing noise caused by dark current of the imaging element, and the control procedure is performed when the operation mode is a mode for acquiring a light image. Controls the light shielding means so that the illumination light is irradiated to the sample only during the exposure period. When the operation mode is a mode for acquiring a dark image, the illumination light to the sample during the exposure period is shielded. It is desirable to control the light shielding means.

  According to the present invention, a light source that emits illumination light, an illumination optical system that guides the illumination light emitted from the light source to irradiate the sample, and a light shielding unit that shields the illumination light guided to the sample. , An imaging apparatus that captures an observation image by a microscope apparatus including an imaging optical system that focuses and images the observation light output from the sample irradiated with the illumination light, and illumination light executed by the imaging apparatus In the control method and the illumination light control program, the light-shielding means of the microscope apparatus is appropriately opened and closed in accordance with the operation mode of the imaging apparatus, so that both comfort during observation and prevention of sample damage such as sample fading can be achieved. it can.

Embodiments of the present invention will be described below with reference to the drawings.
First, a first embodiment according to the present invention will be described with reference to FIGS.
FIG. 1 is a diagram illustrating a configuration of a microscope system.
Here, the microscope system is a configuration in which an electronic camera is connected to a microscope apparatus.

  In FIG. 1, the illumination light emitted from the epi-illumination light source 21 is incident on the trinocular tube unit 5 via the epi-illumination shutter 23 that blocks the illumination light guided to the sample 3, and is further emitted from the epi-illumination light source 21. The sample 3 on the stage 26 is irradiated through the objective lens 27 that is an illumination optical system for guiding the sampled illumination light to irradiate the sample 3. The image of the sample 3 is guided to the eyepiece unit 6 through the objective lens 27 and the trinocular tube unit 5 again, and the observation light output from the sample 3 irradiated with the illumination light is condensed and combined. The image is projected onto an electronic camera 36 that is an imaging device through an imaging lens 100 that images.

The electronic camera 36 and the epi-illumination shutter 23 are electrically connected.
FIG. 2 is a block diagram showing a configuration of an electronic camera used in the microscope system.
A portion surrounded by an alternate long and short dash line in FIG. 2 indicates the configuration of the electronic camera 36.
In FIG. 2, a solid-state image pickup device (CCD: Charge Coupled Device; hereinafter referred to as CCD) 42 picks up a color or black-and-white image and is arranged on the observation optical path together with the photographic eyepiece unit 35 of the microscope apparatus described above. Has been.

  As described with reference to FIG. 1, the illumination light from the epi-illumination light source 21 is incident on the trinocular tube unit 5 through the epi-illumination shutter 23, reflected by the cube unit 30, and further through the objective lens 27. The sample 3 on 26 is irradiated. Then, the image of the sample 3 is again incident on the trinocular tube unit 5 through the objective lens 27, and this time is transmitted through the cube unit 30 and guided to the eyepiece unit 6, and is electronically transmitted through the imaging lens 100. Projected onto the camera 36.

  In the electronic camera 36, a solid-state imaging device (hereinafter simply referred to as a CCD) 42 such as a CCD detects an observation light imaged by the imaging lens 100, thereby obtaining an observation image of the sample 3 projected from the microscope apparatus. Imaging and photoelectric conversion. The CCD 42 is, for example, an interline type that can perform a read operation during charge accumulation.

  A CDS circuit (Correlated Double Sampling) 43 that extracts an image signal component from the output signal of the CCD 42, and an AGC (Automatic Gain) for adjusting the output signal level of the CDS circuit 43 to a predetermined gain value. An amplifier (AMP: AMPLifier; hereinafter referred to as AMP) 44, an OB (Optical Black) clamp circuit 52 that determines a black level of an output signal from the AMP 44, and an OB clamp. An A / D (Analog-Digital) converter 45 that converts an analog signal output from the circuit 52 into a digital signal and an output signal from the A / D converter 45 are connected to control writing and reading to the image memory 46. Image processing such as gradation correction and contour enhancement for the image signal read from the memory controller 55 and the image memory 46 An image signal processing circuit 51 that performs image processing, an image display unit 59 that is a display unit that includes a signal processing circuit that processes the image signal into a displayable form, and a camera built-in storage unit that includes a memory that temporarily stores the image signal. A DRAM (Dynamic Random Access Memory) 56, a compression / decompression circuit 57 for compressing and decompressing the image signal, a recording medium 58 such as a memory card for storing the image signal, setting of the shooting mode, and starting an exposure operation An operation unit 61 including a plurality of switches such as a trigger switch that can generate a trigger signal to be generated, and a timing generator (hereinafter referred to as TG) 53 that generates a driving pulse according to a desired exposure time and driving method of the CCD 42. And a sync generator (hereinafter referred to as SG) 54 for supplying a synchronization signal to the TG 53 and the image signal processing circuit 51. It is configured me.

  In addition, a SUB pulse superimposing circuit 49 for inputting a SUB pulse clamped to the internal voltage of the CCD 42 and a signal indicating the exposure period supplied from the TG 53 are sent to the epi-illumination shutter 23 of the microscope apparatus in accordance with instructions of various operation modes. The shutter control signal generator 90 generates and outputs an open / close control signal. A camera shutter 91 is provided on the front surface of the CCD 42 for projecting an observation image onto the CCD 42 and switching between light shielding.

  Each of these components is electrically connected to a CPU (Central Processing Unit) 60 that is a control means, and the entire electronic camera 36 of the present embodiment is centrally controlled by the CPU 60. . The CCD 42 also has an electronic shutter function so that the exposure time can be controlled.

  Further, a ROM (Read Only Memory) 62 and a RAM (Random Access Memory) 63 are connected to the CPU 60 via a bus, and the CPU 60 reads various programs including the illumination light control program 64 stored in the ROM 62. And execute. In particular, the illumination light control program 64 according to the present invention is executed while the CPU 60 refers to the operation definition table 65 stored in the RAM 63.

Next, the operation of the imaging apparatus that is the electronic camera 36 configured as described above will be described. Here, only the portion related to the present invention is described among the operations performed at the time of photographing.
In the fluorescence observation spectroscopic method, the illumination light irradiated from the epi-illumination light source 21 is dispersed by the fluorescent cube in the cube unit 30 and is irradiated to the sample 3 through the objective lens 27. The sample 3 emits fluorescence that is weaker than the irradiated light, and the emitted light is selectively transmitted through the objective lens 27 by the fluorescent cube in the cube unit 30 through a wavelength component different from that of the above-described spectrum. Projected on.

  In the observation light (image signal) detected by the CCD 42, an image signal component is extracted in the CDS circuit 43, the output signal level is adjusted to a predetermined gain value in the AMP 44, and the black level is adjusted to a predetermined DC voltage in the OB clamp 52. Then, the A / D converter 45 converts it into a digital signal. The image signal converted into the digital signal is temporarily stored in the image memory 46 by the memory controller 55 and is output to the image signal processing circuit 51, and the image signal processing circuit 51 performs image processing such as γ correction and edge enhancement. Is output to the image display unit 59, whereby the observation image of the sample 3 is displayed.

The above is the basic operation when displaying a moving image.
Next, the operation during still image recording will be described.
At the time of still image recording, a still image recording trigger is generated by a still image recording instruction from the operation unit 61, and an image signal is temporarily stored in the image memory 46 as described above. Next, the image signal is read from the image memory 46 and subjected to image processing. The image signal output from the image signal processing circuit 51 is temporarily stored in the DRAM 56, and the compression / decompression circuit 57 performs compression processing on the image signal according to the setting from the operation unit 61. To record on the recording medium 58. If compression setting is not performed from the operation unit 61, still image recording can be performed on the recording medium 58 without compression.

The operation when displaying a moving image will be described with reference to FIGS. Note that the moving image is displayed in the normal state before the still image is captured.
FIG. 3 is a flowchart showing a flow of processing executed by the illumination light control program in moving image display.

FIG. 4 shows the data structure of the action definition table.
First, when the execution of the illumination light control program 64 is started, the CPU 60 refers to the operation definition table 65 (FIG. 4).
In step S31, the epi-illumination shutter 23 that is closed when the operation state of the electronic camera (imaging device) 36 is in an idle state, that is, in a state in which the moving image is not displayed and waiting for acquisition of a still image, In order to set it to “open” corresponding to “displaying”, the epi-illumination shutter 23 is opened so that the illumination light emitted from the epi-illumination light source 21 is irradiated onto the sample 3.

Next, in step S32, a SUB pulse (VSUB) is generated in the i-th (for example, first) frame, and in step S33, the SUB pulse is stopped, and at the same time, charge is accumulated in the CCD 42 by the exposure period signal EXP. To start.
In step S34, charges are accumulated in the CCD 42 until the elapsed time T from the start of accumulation reaches a preset exposure time TEXP (until T = TEXP), and when T = TEXP, step S35 is reached. Then, the transfer pulse TG is generated and the accumulation in the CCD 42 is finished.

  Next, in step S36, output of the i-th (for example, first) frame accumulated in the CCD 42 in steps S33 to S35 is started in the i + 1-th (for example, second) frame period, and at the same time, the output to the memory is started. Writing and image display are started, and an + 1-th frame SUB pulse (VSUB) is generated. Then, in step S37, while continuing to output the i-th frame, writing to the memory and displaying the image, the SUB pulse of the i + 1-th frame is stopped, and at the same time, accumulation of charges in the CCD 42 is started by the exposure period signal EXP. To do.

  In step S38, charge is accumulated in the CCD 42 until the elapsed time T from the start of accumulation of the (i + 1) th frame reaches a preset exposure time TEXP (until T = TEXP), and T = TEXP. In step S39, the output of the i-th frame, the writing to the memory, and the image display are terminated, and the transfer pulse TG for the i + 1-th frame is generated, and the accumulation in the CCD 42 is terminated.

In step S40, for example, it is determined whether or not to stop moving image display based on the presence or absence of an instruction to stop moving image display.
If it is determined in step S40 not to stop (No), i is incremented by 1 (i = i + 1), and step S36 and subsequent steps are repeated for the next frame. On the other hand, if it is determined to stop in step S40 (Yes), in step S41, the epi-illumination shutter 23 is closed so that the illumination light emitted from the epi-illumination light source 21 is not irradiated on the sample 3 and the process is terminated.

By such processing of the illumination light control program, the epi-illumination shutter 23 is controlled so that illumination light is always irradiated to the sample 3 when the operation mode is the moving image display mode (moving image display in FIG. 4).
FIG. 5 is a timing chart for explaining a CCD driving method in moving image display.

  In order from the top in FIG. 5, a frame number, a vertical synchronization signal VD, a substrate applied high voltage pulse VSUB for forcibly discharging charges in the charge storage region to the semiconductor substrate, a transfer pulse TG from the charge storage region to the vertical transfer path, and imaging. It shows the charge accumulation state of the element (CCD), the imaging element (CCD) output, the memory operation state, the image display state, the exposure period signal EXP, and the microscope shutter control signal.

  During one frame (between the vertical synchronization signal VD and the next vertical synchronization signal VD), the period superimposed on the SUB pulse (VSUB) is a charge for forcibly discharging the charge accumulated in the charge accumulation region to the semiconductor substrate. Is not stored in the CCD 42. On the other hand, charges are accumulated in the CCD 42 during the period when the SUB pulse is not input to the CCD 42 (CCD accumulation 1 in FIG. 5). That is, the exposure period is a period during which no SUB pulse is input. Note that the exposure period can be changed by changing the period in which the pulse is superimposed on the VSUB.

  The signal charge accumulated in the CCD 42 is read out to a vertical transfer path (not shown) in the CCD 42 by a charge transfer pulse TG. The signal charge transferred through the vertical transfer path in the next frame (the period of the next vertical synchronization signal VD) is transferred to a horizontal transfer path (not shown), read out from the horizontal transfer path, transferred in the direction of AMP 44, and passed through the charge detector. Finally, the pixels are sequentially output one by one by the readout amplifier (CCD output 1 in FIG. 5).

On the other hand, the CCD 42 accumulates the signal charges of the next frame in the charge accumulation region during a predetermined exposure period while reading out the signal charges.
In this way, the CCD 42 performs an exposure operation for accumulating charge every frame. That is, a signal representing the exposure period of each frame is output from the transfer pulse TG.

Here, the shutter control signal generation unit 90 is set by the CPU 60 that the operation mode is during the moving image display period. In the operation definition table 65 (see FIG. 4), the epi-illumination shutter 23 is set during the moving image display period. The action is defined so that it is always open.
Accordingly, during the moving image display period, the shutter control signal generation unit 90 outputs a signal that opens the epi-illumination shutter 23 to the epi-illumination shutter 23. In FIG. 5, the output of the microscope shutter control signal is at the Hi level and the signal level at which the epi-illumination shutter 23 is opened.

Next, the operation at the time of taking a still image will be described with reference to FIG.
FIG. 6 is a timing chart for explaining a CCD driving method in capturing a still image.
In order from the top in FIG. 6, the frame number, the vertical synchronization signal VD, the substrate applied high voltage pulse VSUB for forcibly discharging the charge storage region charge to the semiconductor substrate, the transfer pulse TG from the charge storage region to the vertical transfer path, and imaging It shows the charge accumulation state of the element (CCD), the image pickup element (CCD) output, the memory operation state, the image display state, the still image recording state, the exposure period signal EXP, and the microscope shutter control signal.

The still image capturing operation is started by a still image recording trigger from the operation unit 61 (still image recording trigger in FIG. 6).
Until the still image recording trigger is given, the electronic camera 36 is performing the moving image display operation, so that the epi-illumination shutter 23 is open. When a still image recording trigger is given, exposure for a still image is performed in the next vertical synchronizing signal VD period.

  The observation light is accumulated in the CCD 42 for a predetermined exposure period (CCD accumulation 3 in FIG. 6). Next, the charges accumulated by the charge transfer pulse TG are read out to the vertical transfer path, and are output from the CCD 42 (CCD output 3 in FIG. 6), and temporarily stored in the image memory 46 (FIG. 6). Memory "3 write"). In the vertical synchronization signal VD period, the image signal is read from the image memory 46, subjected to image processing, and recorded on the recording medium 58.

  Here, during the still image recording operation, the operation definition table 65 defines that the epi-illumination shutter 23 is opened only during the still image exposure period. Therefore, the microscope shutter control signal during the still image recording operation outputs a signal that opens the epi-illumination shutter 23 only during the exposure period for the still image, and for the period when the accumulated charges are discharged by the SUB pulse and for the still image. A signal that closes the epi-illumination shutter 23 is output during a period in which the accumulated charge is read.

In FIG. 6, the output of the microscope shutter control signal is a signal level for opening the epi-illumination shutter 23 when the Hi level is high, and the signal level for closing the epi-illumination shutter 23 when the microscope shutter control signal is the Lo level.
By such processing, the epi-illumination shutter 23 is controlled so that the illumination light is irradiated to the sample 3 only during the exposure period when the operation mode is the still image display mode.

  Since the basic operation described with reference to FIGS. 3 to 6 does not open or close the epi-illumination shutter 23 every frame during the moving image display period and no shutter operation sound is generated, the user is not required. Observation can be performed comfortably without listening to the sound. During the still image recording operation, the sample 3 is irradiated with excitation light only for a necessary period. 3 damage prevention can be achieved.

Next, an operation at the time of intermittent shooting in which the electronic camera 36 operating as described above records the change with time of the sample 3 will be described.
The moving image display period is the same as described above until the still image recording trigger is given.
FIG. 7 is a flowchart showing a flow of processing executed by the illumination light control program in intermittent shooting.

First, in step S61, the epi-illumination shutter 23 is closed, and in step S62, initialization is performed by substituting 0 into a counter FRM that counts the number of frames taken during intermittent shooting.
In step S63, it is determined whether there is an instruction for intermittent shooting (ON). If it is determined that there is an instruction (Yes), a shooting interval timer (intermittent timer) for intermittent shooting is determined in step S64. Initialization is performed by substituting 0 into TINTVL, and in step S65, counting up of the photographing interval timer (intermittent timer) TINTVL is started.

  Next, in step S66, a SUB pulse (VSUB) is generated, and in step S67, the epi-illumination shutter 23 is opened. In step S68, the SUB pulse is stopped, and at the same time, charges are accumulated in the CCD 42 by the exposure period signal EXP. Start.

  In step S69, charge is accumulated in the CCD 42 until the elapsed time T from the start of accumulation reaches a preset exposure time TEXP (until T = TEXP), and when T = TEXP, step S70 is reached. Then, the transfer pulse TG is generated and the accumulation in the CCD 42 is finished.

  Next, in step S71, the epi-illumination shutter 23 is closed, and in step S72, output of the electric charge accumulated in the CCD 42 in steps S68 to S70 is started, and writing into the memory and image display are started. In step S73, Record a still image.

Since the still image is recorded in step S73, the intermittent shooting frame number FRM is incremented by 1 in step S74 (FRM = FRM + 1), and it is determined again in step S75 whether or not the intermittent shooting is ON.
If it is determined in step S75 that intermittent shooting is ON (Yes), it is determined in step S76 whether the number of intermittent shooting frames FRM has reached a preset number of frames FINTVL (FRM = FINTVL). ) Is determined (Yes), the process is terminated (Yes), but if not reached (No), the shooting interval timer (intermittent timer) TINTVL is reached in step S77 until the predetermined interval TINTVL is reached (TMINTVL). = TINTVL) and wait (Yes), repeat step S64 and subsequent steps.

FIG. 8 is a timing chart for explaining a CCD driving method in intermittent shooting.
In FIG. 8, in order from the top, the frame number, the vertical synchronization signal VD, the substrate applied high voltage pulse VSUB for forcibly discharging the charges in the charge storage region to the semiconductor substrate, the transfer pulse TG from the charge storage region to the vertical transfer path, and imaging It shows the charge accumulation state of the element (CCD), the image pickup element (CCD) output, the memory operation state, the image display state, the still image recording state, the exposure period signal EXP, and the microscope shutter control signal.

The number of intermittent shooting and the shooting interval are set in advance from the operation unit 61, and the moving image is not displayed during the shooting standby period.
When a still image recording trigger is given from the operation unit 61, charges are accumulated in the CCD 42 for a predetermined accumulation period as in the still image recording operation (CCD accumulation 3 in FIG. 8). At this time, the shutter control signal generator 90 outputs a Hi level signal only during the exposure period for recording a still image.

Next, the accumulated charge is output from the CCD 42 in the next vertical synchronizing signal VD period (CCD output 3 in FIG. 8) and temporarily stored in the image memory 46. During the vertical synchronization signal VD, the shutter control signal generator 90 outputs a Lo level signal so as to close the epi-illumination shutter 23.
Further, in the next vertical synchronizing signal VD period, the image signal output from the image memory 46 is subjected to image processing in the image signal processing circuit 51 (image processing 3 in FIG. 8), and is applied to the recording medium 58 in the same manner as the still image recording operation. To be recorded.

Since the moving image display is not performed during the imaging standby period until the next imaging including the image processing period and the recording period to the recording medium 58, the shutter control signal generation unit 90 outputs the Lo level signal to close the epi-illumination shutter 23. Output.
When a preset shooting interval elapses, exposure for the next still image recording is performed, and the shutter control signal generation unit 90 outputs a Hi level signal to open the epi-illumination shutter 23 during the exposure period.

The signal charge accumulated at this time (CCD accumulation 8 in FIG. 8) is output from the CCD 42 (CCD output 8 in FIG. 8) in the next vertical synchronization signal VD period, temporarily stored in the image memory 46, and further, the next vertical synchronization. Recording is performed on the recording medium 58 during the signal VD period.
The above operation is repeated for a predetermined number of shots. When recording for a predetermined number of images is completed, exposure for moving image display is started again (CCD accumulation 13 in FIG. 8).

At the same time, the shutter control signal generator 90 outputs a Hi level signal so that the epi-illumination shutter 23 is always opened again from this period.
As described above, when it is set not to display a moving image during standby during intermittent shooting, the epi-illumination shutter 23 is closed while the moving image is not displayed before and after the intermittent shooting operation, and a still image is displayed during the intermittent shooting operation. Since the epi-illumination shutter 23 is opened only during the exposure period for recording, the illumination light is irradiated on the sample 3 only for a necessary period at the time of recording a still image, and before and after displaying a moving image while suppressing damage such as discoloration of the sample 3 It is possible to observe comfortably without hearing troublesome shutter operation sound.

  The above is the operation when the moving image is not displayed during intermittent shooting. For example, if “automatic exposure” is set during shooting standby from the operation unit 61, automatic shooting for the next shooting during shooting standby is performed. It is also possible to perform exposure setting. In this case, the CPU 60 temporarily puts a moving image in the image acquisition state during standby for shooting, and during that period, the shutter control signal generator 90 opens the epi-illumination shutter 23. When the automatic exposure setting is completed, the shutter control signal generation unit 90 may perform an operation of closing the epi-illumination shutter 23 by canceling the moving image display state.

Next, a second embodiment according to the present invention will be described with reference to FIGS.
The second embodiment is an embodiment of an operation at the time of dark noise suppression processing that generally depends on the exposure time and temperature of an image sensor (CCD). Note that the operation during the moving image display is the same as that in the first embodiment, and a description thereof will be omitted.

FIG. 9 is a diagram for explaining dark noise suppression processing.
In FIG. 9, an image memory 46 includes an image memory 1 (46-1) and an image memory 2 (46-2), and an image signal processing circuit 51 includes a calculation unit 80, a gradation correction unit 81, and a contour enhancement unit. 82.

  The image memory 1 (46-1) stores a bright image, and the image memory 2 (46-2) stores a dark image. Then, the calculation unit 80 subtracts the dark image stored in the image memory 2 (46-2) from the bright image stored in the image memory 1 (46-1), resulting from the dark current of the CCD 42. Suppresses noise. The gradation correction unit 81 corrects the gradation of the image data subtracted by the calculation unit 80, and the contour emphasizing unit 82 further performs a process of enhancing the contour.

FIG. 10 is a flowchart showing a flow of processing executed by the illumination light control program in dark noise suppression processing.
First, in step S81, from the state where the epi-illumination shutter 23 is closed, a SUB pulse (VSUB) is generated in step S82, and the epi-illumination shutter 23 is opened in step S83.

Next, in step S84, the SUB pulse is stopped, and at the same time, the accumulation of electric charges in the CCD 42 for the bright image is started by the exposure period signal EXP.
In step S85, until the elapsed time T from the start of accumulation reaches a preset exposure time TEXP (until T = TEXP), the charge is accumulated in the CCD 42 for the bright image, and T = TEXP. In step S86, a transfer pulse TG is generated, and the accumulation of the bright image in the CCD 42 is terminated.

  Next, in step S87, the epi-illumination shutter 23 is closed, and in step S88, output of electric charges and writing to the memory for the bright image accumulated in the CCD 42 in steps S84 to S86 are started, and for the dark image. SUB pulse (VSUB) is generated.

In step S89, the output of the bright image and the writing to the memory are terminated, and the SUB pulse is stopped. At the same time, the accumulation of electric charges in the CCD 42 for the dark image is started by the exposure period signal EXP.
Next, in step S90, until the elapsed time T from the start of accumulation reaches a preset exposure time TEXP (until T = TEXP), the charge is accumulated in the CCD 42 for the implicit image, and T = If it becomes TEXP, a transfer pulse TG is generated in step S91, and the accumulation of the dark image in the CCD 42 is terminated.

In step S92, electric charges are output and written to the memory for the dark image stored in the CCD 42 in steps S89 to S91.
Next, in step S93, dark noise suppression calculation processing, that is, processing for subtracting the dark image from the bright image, gradation correction processing, contour enhancement processing, and the like are performed. In step S94, the still image is processed. Record.

FIG. 11 is a timing chart for explaining a CCD driving method in dark noise suppression processing.
In order from the top in FIG. 11, a frame number, a vertical synchronization signal VD, a substrate applied high voltage pulse VSUB for forcibly discharging charges in the charge storage region to the semiconductor substrate, a transfer pulse TG from the charge storage region to the vertical transfer path, imaging Device (CCD) charge accumulation state, imaging device (CCD) output, memory operation state, dark noise suppression processing state, image display state, still image recording state, camera shutter control signal, exposure period signal EXP, microscope shutter control signal Is shown. Note that the timing chart of FIG. 11 shows the operation during long exposure during still image recording.

When performing dark noise suppression processing, first, a photographed image (light image) is actually exposed with the camera shutter 91 opened. Next, the camera shutter 91 is closed, and the image (dark image) when the CCD 42 is shielded from light is exposed for the same time as the light image.
Then, dark noise suppression processing is performed by subtracting the dark image from the bright image.

When dark noise suppression is set from the operation unit 61, when a still image recording trigger is given, exposure of a bright image for still image recording is started with the next vertical synchronization signal VD.
In the case of long exposure, the period from when the SUB pulse (VSUB) stops to when the charge transfer pulse TG is input is the exposure period. In FIG. 11, the 2 vertical synchronization signal VD period is the exposure period. Is accumulated (CCD accumulation 2 in FIG. 11).

At this time, the camera shutter 91 is in the open state, and the epi-illumination shutter 23 is in the open state only during the exposure period of the bright image.
When the charge transfer pulse TG is input, the exposure period of the bright image ends, the CPU 60 closes the camera shutter 91, and the shutter control signal generator 90 outputs a Lo level signal to close the epi-illumination shutter 23.

In FIG. 11, the camera shutter 91 is opened when the camera shutter control signal Hi level, and the camera shutter 91 is shielded when the camera shutter control signal is Lo level.
When the charge transfer pulse TG is inputted, the accumulated charge is outputted from the CCD 42 by the next vertical synchronizing signal VD (CCD output 2 in FIG. 11). The output image from the CCD 42 is temporarily stored in the image memory 1 (46-1).

At the same time when the bright image is output from the CCD 42, the CCD 42 is shielded from light so that exposure of the dark image is started in the CCD 42 and charge is accumulated until the charge transfer pulse TG is input (CCD accumulation in FIG. 11). 3).
At this time, the CCD 42 only needs to be able to store the light-shielded image, so the shutter control signal generation unit 90 outputs the Lo level signal to the epi-illumination shutter 23 even during the exposure period of the dark image so that the epi-illumination shutter 23 is closed.

When the charge transfer pulse TG is input to the CCD 42, the CCD 42 outputs the accumulated charge by the next vertical synchronization signal VD (CCD output 3 in FIG. 11). This output signal is temporarily stored in the image memory 2 (46-2).
When the bright image and the dark image are stored in the image memory 46, dark noise suppression processing is performed in the next vertical synchronization signal VD period. The memory controller 55 simultaneously reads out the image signal from the image memory 1 (46-1) and the image memory 2 (46-2), and the arithmetic unit 80 subtracts the dark image from the bright image (dark noise suppression in FIG. 11). Process 2-3 (2 minus 3)).

The subtraction result in the calculation unit 80 is subjected to other image processing, such as gradation correction processing and contour enhancement processing, in the image signal processing unit 51, and is output to the image display unit 59 and is temporarily stored in the DRAM 56 and is temporarily stored in the recording medium 58. Recorded as a still image.
When the still image is recorded, the CPU 60 opens the camera shutter 91 to resume moving image display, and the shutter control signal generation unit 90 outputs a Hi level signal to the microscope control unit 41 to control the epi-illumination shutter 23 to the open state. To do.

  When dark noise suppression processing involving opening and closing of the camera shutter 91 is performed by the above operation, the epi-illumination shutter 23 is always open during the moving image display period, and the epi-illumination shutter 23 is activated during the dark image exposure period during still image recording. Since the epi-illumination shutter 23 is opened only during the exposure period of the bright-time image, the illumination light is irradiated on the sample 3 only for a necessary period at the time of still image recording, and before and after the moving image while suppressing damage such as fading of the sample 3 During display, it is possible to observe comfortably without hearing a troublesome shutter operation sound. That is, when the operation mode is a mode for acquiring a light image, the epi-illumination shutter 23 is controlled so that illumination light is irradiated on the sample 3 only during the exposure period, and the operation mode is a mode for acquiring a dark image. In this case, the epi-illumination shutter 23 is controlled so as to block the illumination light to the sample 3 during the exposure period.

  As described above, the first embodiment and the second embodiment to which the present invention is applied have been described with reference to FIGS. 1 to 11, and the epi-illumination light source 21 and the epi-illumination shutter 23 described in these embodiments. Instead of the imaging apparatus (electronic camera 36), the microscope apparatus using the transmission illumination light source 13 and the transmission shutter 161 is used depending on the microscopy method of the microscope apparatus, as in the microscope system in the first modification shown in FIG. And may be combined.

  Further, like the electronic camera 37 used in the microscope system in the second modification shown in FIG. 13, some functions of the electronic camera 36 described above, that is, the image memory 46, the image signal processing circuit 51, and the memory controller 55. The DRAM 56, the compression / decompression circuit 57, the recording medium 58, the image display unit 59, the CPU 60, the operation unit 61, the ROM 62, and the RAM 63 may be provided in an external computer (PC) 170 via the external I / F 160.

  As described above, each embodiment and modification of the present invention have been described with reference to the drawings. However, an imaging apparatus to which the present invention is applied can be used as long as the functions thereof are executed. The system may be a single device, a system composed of a plurality of devices or an integrated device, or a system that performs processing via a network such as a LAN or WAN. Needless to say.

  It can also be realized by a system comprising a CPU, ROM or RAM memory connected to the bus, input device, output device, external recording device, medium driving device, portable recording medium, and network connection device. That is, a ROM or RAM memory, an external recording device, and a portable recording medium that record the program code of the software that realizes the system of each of the above-described embodiments is supplied to the imaging device, and the computer of the imaging device performs the program. Needless to say, this can also be achieved by reading and executing the code.

In this case, the program code itself read from the portable recording medium or the like realizes the novel function of the present invention, and the portable recording medium or the like on which the program code is recorded constitutes the present invention. .
Examples of portable recording media for supplying program codes include flexible disks, hard disks, optical disks, magneto-optical disks, CD-ROMs, CD-Rs, DVD-ROMs, DVD-RAMs, magnetic tapes, and non-volatile memories. Various recording media recorded through a network connection device (in other words, a communication line) such as a card, a ROM card, electronic mail or personal computer communication can be used.

  In addition, the functions of the above-described embodiments are realized by executing the program code read out on the memory by the computer, and the OS running on the computer is actually executed based on the instruction of the program code. The functions of the above-described embodiments are also realized by performing part or all of the process.

  Furthermore, a program code read from a portable recording medium or a program (data) provided by a program (data) provider is provided in a function expansion board inserted into a computer or a function expansion unit connected to a computer. The CPU of the function expansion board or function expansion unit performs part or all of the actual processing based on the instruction of the program code, and the functions of the above-described embodiments are also performed by the processing. Can be realized.

  That is, the present invention is not limited to the above-described embodiments and the like, and can take various configurations or shapes without departing from the gist of the present invention.

It is a figure which shows the structure of a microscope system. It is a block diagram which shows the structure of the electronic camera used for a microscope system. It is a flowchart which shows the flow of the process which an illumination light control program performs in a moving image display. It is a figure which shows the data structure of an action definition table. 6 is a timing chart for explaining a CCD driving method in moving image display. 4 is a timing chart for explaining a method of driving a CCD in capturing a still image. It is a flowchart which shows the flow of the process which an illumination light control program performs in intermittent imaging | photography. 6 is a timing chart for explaining a CCD driving method in intermittent shooting. It is a figure for demonstrating the suppression process of dark noise. It is a flowchart which shows the flow of the process which an illumination light control program performs in the suppression process of dark noise. 6 is a timing chart for explaining a CCD driving method in dark noise suppression processing; It is a figure which shows the structure of the microscope system in a 1st modification. It is a block diagram which shows the structure of the electronic camera used for the microscope system in a 2nd modification.

Explanation of symbols

DESCRIPTION OF SYMBOLS 3 Sample 5 Trinocular tube unit 6 Eyepiece unit 13 Transmitted illumination light source 21 Epi-illumination light source 23 Epi-illumination shutter 26 Stage 27 Objective lens 30 Cube unit 36 Electronic camera 37 Electronic camera 42 Solid-state image sensor (CCD)
43 CDS circuit 44 Amplifier (AMP)
45 A / D converter 46 Image memory 46-1 Image memory 1
46-2 Image memory 2
49 SUB pulse superposition circuit 51 Image signal processing circuit 52 OB clamp circuit 53 Timing generator (TG)
54 Sync Generator (SG)
55 Memory controller 56 DRAM
57 Compression / Expansion Circuit 58 Recording Medium 59 Image Display Unit 60 CPU
61 Operation section 62 ROM
63 RAM
64 Illumination light control program 65 Operation definition table 80 Calculation unit 81 Gradation correction unit 82 Outline emphasis unit 90 Shutter control signal generation unit 91 Camera shutter 100 Imaging lens 160 External I / F
161 Transmission shutter 170 Computer (PC)

Claims (11)

A light source that emits illumination light, an illumination optical system that guides the illumination light emitted from the light source to irradiate the sample, a light shielding unit that shields the illumination light guided to the sample, and the illumination light irradiates An imaging device that captures an observation image by a microscope apparatus including an imaging optical system that focuses and images the observation light output from the sample,
An image sensor for detecting observation light imaged by the imaging optical system;
Display means for displaying an image of the sample based on the observation light detected by the imaging device;
Control means for controlling the light shielding means corresponding to the operation mode displayed by the display means;
An imaging apparatus comprising:   The control means controls the light shielding means so that illumination light is always irradiated to the sample when the operation mode is the moving image display mode, and during the exposure period when the operation mode is the still image display mode. The imaging apparatus according to claim 1, wherein the light shielding unit is controlled so that only the sample is irradiated with illumination light. Further comprising noise suppression means for suppressing noise caused by dark current of the image sensor;
The control means controls the light shielding means so that illumination light is irradiated to the sample only during an exposure period when the operation mode is a mode for acquiring a light image, and the operation mode is a dark image. The imaging apparatus according to claim 1, wherein in the acquisition mode, the light shielding unit is controlled so as to shield illumination light to the sample during an exposure period. The light source is an epi-illumination light source;
4. The epi-illumination shutter according to claim 1, wherein the light shielding means is an epi-illumination shutter that illuminates the sample with the illumination light when opened and shields the illumination light when closed. The imaging device described. The light source is a transmitted illumination light source;
4. The transmission shutter according to claim 1, wherein the light shielding unit is a transmissive shutter that irradiates the sample with the illumination light when opened and blocks the illumination light when closed. 5. The imaging device described. A light source that emits illumination light, an illumination optical system that guides the illumination light emitted from the light source to irradiate the sample, a light shielding unit that shields the illumination light guided to the sample, and the illumination light irradiates An illumination light control method executed by an imaging device that captures an observation image by a microscope device that includes an imaging optical system that focuses and images the observation light output from the sample,
Detecting observation light imaged by the imaging optical system;
Based on the detected observation light, when displaying an image of the sample, the light shielding means is controlled corresponding to an operation mode,
The illumination light control method characterized by the above-mentioned.   The control controls the light shielding means so that illumination light is always irradiated to the sample when the operation mode is the moving image display mode, and only during the exposure period when the operation mode is the still image display mode. The illumination light control method according to claim 6, wherein the light shielding unit is controlled so that illumination light is irradiated onto the sample. Furthermore, the noise caused by the dark current of the image sensor is suppressed,
In the control, when the operation mode is a mode for acquiring a light-time image, the light shielding unit is controlled so that illumination light is irradiated on the sample only during an exposure period, and the operation mode acquires a dark-time image. 8. The illumination light control method according to claim 6, wherein the light shielding unit is controlled so that illumination light to the sample during an exposure period is shielded during the exposure mode. A light source that emits illumination light, an illumination optical system that guides the illumination light emitted from the light source to irradiate the sample, a light shielding unit that shields the illumination light guided to the sample, and the illumination light irradiates An illumination light control program for causing an imaging device that captures an observation image by a microscope device provided with an imaging optical system that focuses and images the observation light output from the sample,
A procedure for detecting observation light imaged by the imaging optical system;
Based on the detected observation light, when displaying an image of the sample, a procedure for controlling the light shielding means corresponding to an operation mode;
A computer-executable illumination light control program for executing the program.   The controlling procedure controls the light shielding means so that illumination light is always irradiated to the sample when the operation mode is the moving image display mode, and the exposure period when the operation mode is the still image display mode. The illumination light control program according to claim 9, wherein the light shielding unit is controlled so that illumination light is irradiated only on the sample. Further comprising a procedure for suppressing noise caused by dark current of the image sensor;
In the control procedure, when the operation mode is a mode for acquiring a light image, the light shielding unit is controlled so that illumination light is irradiated to the sample only during an exposure period, and the operation mode is a dark image. 11. The illumination light control program according to claim 9, wherein the light shielding unit is controlled so that illumination light to the sample during an exposure period is shielded in a mode for acquiring the light.

Images